SPSFAM ERP

Procedure: 1. Place a 250 mL Nalgene polypropylene bottle on the balance and tare it to zero. 2. Dispense a minimum of 7.5 g sample 5 into the container and record the weight. 3. Add 100 mL of diluant 5 mg/mL IS to the container using an automatic pipet or calibrated bottle top dispenser and securely fasten the bottle with a screw cap lid. 4. Perform a manual shake of the bottle and inspect if the contents are visually dissolving in solution, and place these samples on the shaker for a minimum of 15 minutes at a rate of 110-130 shakes per minute. 5. For samples not visually dissolving, microwave each individual bottle for 25 seconds and then place on the shaker for a minimum of 15 minutes at a rate of 110-130 shakes per minute. Representative Food Matrices requiring the Microwave Step (not an exclusive list): Gelatin, pectin, guar gum etc. Peanut butter, ground nuts Cottage or cream cheese Potato or corn chips Date or almond paste Dried fruit Butter, margarine mayonnaise, oil Frosting Chocolate bar Cheese 6. Post shaking, let solids settle out of solution to the bottom and fat rise to the surface for not less than 10 minutes. 7. If after 10 minutes the samples do not have a clear region to acquire an aliquot, centrifuge sample in a disposable centrifuge tube for 5 minutes at 3000 RPM 6 . 8. Pipette 2 mL of aqueous extract into a 2 dram vial. 9. Dispense 3 mL of acetonitrile into each 2 dram vial and briefly vortex mix. 10. Let mixture sit for a minimum of 15 minutes to allow the precipitation of proteins to the bottom of the tube and oleaginous materials rise to the surface 7 . 5 For samples containing >50% estimated sugar content (i.e. marbits), weigh samples accordingly, (eg. less). For samples containing >70% estimated water (i.e. yogurt and beverages), weigh 20 g of sample. For all other matrices, inclusive of maltodextrin or soluble corn fiber samples, weigh a minimum of 7.5g of sample. 6 Some RTE cereals, feed samples, fiber ingredients, and infant formula will require centrifugation. Infant formula and milk were found to require a minimum of 10 min centrifugation. 7 If dilution is required for quantitation of any individual sugar, an aliquot from this solution may be diluted with 60:40 acetonitrile: water using positive displacement pipettes.